Supplementary Figure 4: NOTCH1 drives autonomous and non-autonomous cell-cycle arrest in part through TGF-β.
From: NOTCH1 mediates a switch between two distinct secretomes during senescence
(a) Left: proliferation analysis of TRE-N1ICD IMR90 cells treated with or without doxycycline with or without the TGF-β receptor antagonist GW788388. Right: The proliferative ability of mRFP IMR90 cells was analysed during co-culture with TRE-N1ICD IMR90 cells treated with or without doxycycline and GW788388; Values are mean ± s.e.m., representative of three independent experiments with similar results. (b) Left: proliferation analysis of TRE-N1ICD IMR90 cells with dnSMAD4-mVenus or vector control treated with or without doxycycline. Middle: The proliferative ability of mRFP IMR90 cells was analysed during co-culture with TRE-N1ICD IMR90 cells with dnSMAD4-mVenus or vector control treated with or without doxycycline; Values are mean ± s.e.m., representative of three independent experiments with similar results. Right: Immunofluorescence for TGFBI in IMR90 cells with dnSMAD4-mVenus or vector control treated with 10 ng ml−1 TGF-β1 for 8 h. Scale bar 30 μm. (c) IMR90 cells were subject to short-term proliferation analysis with or without the indicated concentrations of recombinant TGF-β1, 2 or 3; values are mean ± s.e.m.; representative of three independent experiments with similar results. (d) Representative immunofluorescence of SMAD2/3 after 24 h treatment with 10 ng ml−1 TGF-β1, 2, 3 or vehicle. Scale bar 50 μm.
